ARID4B在牛磺酸调节成肌细胞C2C12蛋白质合成中的作用  

Role of ARID4B in stimulation of taurine on protein synthesis in C2C12 myoblast cells

作　　者：高学军[1] 王璐璐 祁昊 王哲 甄贞[1] GAO Xuejun;WANG Lulu;QI Hao;WANG Zhe;ZHEN Zhen(School of Life Science,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2021年第1期46-53,共8页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(31671473)。

摘　　要：肌肉细胞蛋白质合成能力与畜禽产肉量性状有关,试验旨在探究转录因子AT富集区4B(AT-rich interaction domain 4B,ARID4B)对牛磺酸(Taurine,Tau)调节成肌细胞C2C12蛋白质合成的影响。向体外培养C2C12细胞培养液中分别添加0、60、120、180和240μmol·L^(-1) Tau,采用BCA(Bicinchoniic acid)蛋白定量试剂盒和SUnSET法分别检测细胞总蛋白质含量和蛋白质合成速率,采用Western blot检测细胞中蛋白质合成相关信号通路分子mTOR磷酸化变化。进一步检测添加最适浓度Tau同时转染siRNA敲低ARID4B对mTOR磷酸化和mTOR mRNA水平的影响。结果表明,Tau以剂量依赖性方式促进C2C12细胞蛋白质合成。当Tau浓度为120μmol·L^(-1)时,蛋白质合成总量、蛋白质合成速率和mTOR磷酸化水平达到峰值,后逐渐下降。Tau显著促进ARID4B蛋白表达,敲低ARID4B后,细胞蛋白质合成以及mTOR磷酸化水平显著降低,敲低ARID4B也显著抑制Tau对蛋白质合成、mTOR磷酸化以及mTOR mRNA表达促进作用。综上,ARID4B通过正向调节mTOR mRNA表达促进mTOR磷酸化,调节C2C12细胞中蛋白质合成。The ability of protein synthesis in muscle cells is related to meat yield traits of livestock and poultry,and the aim of this study was to explore the effects of taurine(Tau)on protein synthesis ability in myoblast C2C12 and the role of AT-rich interaction domain 4B(ARID4B)in this regulatory process.In this experiment,cells were treated with 0,60,120,180 and 240μmol·L^(-1) Tau.Total protein content and protein synthesis rate were measured using the BCA protein quantitative assay kit and the SUnSET method,respectively.The phosphorylation of mTOR,which was a major signaling molecule associated with protein synthesis,was also detected by Western blot.The effects of knockdown of ARID4B in the presence of Tau on mTOR phosphorylation and mTOR mRNA expression were further determined by Western blot and qRT-PCR analysis,respectively.The results showed that Tau promoted protein synthesis in C2C12 cells in a dose-dependent manner.When Tau concentration was at 120μmol·L^(-1),the protein synthesis ability and mTOR phosphorylation level peaked,and then both gradually decreased with the increase of Tau concentration.Tau significantly promoted ARID4B protein expression.After knocking down ARID4B,cellular protein synthesis and mTOR phosphorylation levels were significantly reduced.Knockdown of ARID4B also significantly inhibited the promotion of protein synthesis,mTOR phosphorylation and mTOR mRNA expression by Tau.In summary,ARID4B positively regulated mTOR mRNA expression and promoted mTOR phosphorylation to regulate protein synthesis in C2C12 cells.

关 键 词：C2C12 牛磺酸 MTOR 蛋白质合成 ARID4B 

分 类 号：S823[农业科学—畜牧学]